CXCR4/CXCL12、CD44和CD147标记的垂体腺瘤细胞在其侵袭性生物行为中的表达意义
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摘要
目的研究CXCR4、CXCL12、CD44、CD147在侵袭性垂体腺瘤和非侵袭性垂体腺瘤中的表达水平,通过比较分析其与垂体腺瘤侵袭性的相关性及其可能机制。
材料与方法收集2009年1月-4月在我院神经外科手术切除的35例新鲜垂体腺瘤标本,制成单细胞悬液,采用流式细胞仪检测CXCR、CD44、CD147的表达水平；同时将检测的细胞悬液,在荧光显微镜下观察细胞的荧光标记情况拍照记录；并将与其对应的垂体腺瘤石蜡标本,应用免疫组化的方法,研究CXCL12/CXCR4、CD44和CD147的表达情况；采用SPSS17.0统计处理。
结果依据内分泌临床表现、影像学检查、术中所见和病理诊断结果,将垂体腺瘤(35例)分为21例侵袭性垂体腺瘤和14例非侵袭性垂体腺瘤两个实验组。比较两组间肿瘤标记物的表达水平,流式细胞仪检测结果中CXCR4、CD44、CD147的P值分别为0.002、0.221和0.178,免疫组化结果中CXCR4、CXCL12、CD44、CD147的P值分别为0.04、0.01、0.516和0.855；垂体腺瘤中CXCR4/CXCL12的表达水平水平明显高于正常垂体；荧光显微镜观察显示肿瘤细胞部分表达所标记的肿瘤标记物。侵袭鞍底的垂体腺瘤CXCR4表达率明显高于侵袭海绵窦组(mean分别为58.3%和23.6%,P<0.01)。CXCR4/CXCL12、CD44、CD147与患者性别、年龄、肿瘤大小及功能无明显关系(P>0.05)。
结论流式细胞仪检测肿瘤标记物在垂体腺瘤细胞中的表达率上要比免疫组化准确、敏感；在侵袭性垂体腺瘤中CXCR4/CXCL12的表达水平显著升高,CD44、CD147的表达水平无明显差异。CXCR4/CXCL12可能通过ERK1/2MAP激酶级联反应和Pyk2的活化,在垂体腺瘤的侵袭性发生过程中起作用,可作为研究垂体腺瘤不良生物学行为的新的潜在标记物。
Objective Detect the expression levels of CXCR4, CXCL12, CD44, CD147in invasive and non-invasive pituitary adenomas, and comparatively analyze the relevance of these proteins and the invasiveness of pituitary adenoma and the probable mechanisms.
Materials and Methods Collected the fresh specimens of surgical resection of35pituitary adenoms of our neurosurgery department from January to April,2009; made them to single cell suspension, and detected the expression of CXCR4, CD44, CD147by flow cytometry; at the same time, put the suspension under fluorescence microscope, observed the fluorescent-labeled cells and took photographed records; used the immunohistochemical methods to research the expression of CXCR4, CXCL12, CD44, CD147in the corresponding paraffin specimens of pituitary adenoma, and photographed them; used the SPSS17.0for statistical analysis, and took P<0.05as statistically significant standard.
Results Based on clinical manifestations, imaging examinations, surgery and pathological diagnosis, the pituitary adenomas (35cases) were divided into invasive pituitary adenomas (21cases) and non-invasive pituitary adenomas (14cases) compairing Comparing the expression levels of these two groups, the P values of CXCR4, CD44and CD147were0.002,0.221and0.178respectively in the results of flow cytometry; the P values of CXCR4, CXCL12, CD44, CD147in immunohistochemistry method were0.04,0.01,0.516and0.855respectively; the expression levels of CXCR4/CXCL12in pituitary adenomas were higher than the levels in normal pituitary; fluorescence microscope showed that part of pituitary adenomas expressd the labelled tumor markers. The expression of CXCR4in pituitary adenoma which invaded buttom of sella turcica was significantly higher than that cavernous sinus invaded group(means were58.3%and23.6%, P<0.01). CXCR4/CXCL12, CD44, CD147had no significant relationship with patient gender, age, tumor size and function(P>0.05).
Conclusion Flow cytometry is more accurate and sensitive in the detection of expression rate of tumor markers in tumor cells than immunohistochemistry.In invasive pituitary pituitary adenomas the expressions of CXCR4/CXCL12are significant increased, the expressions of CD44, CD147has no significant difference. CXCR4/CXCL12play an important role in the process of invasiveness of pituitary adenoma, possibly though the ERK1/2MAP kinase cascade and Pyk2activation and can be a new potential marker for the study of the abnormal biological behaviors of pituitary adenoma.
材料与方法收集2009年1月-4月在我院神经外科手术切除的35例新鲜垂体腺瘤标本,制成单细胞悬液,采用流式细胞仪检测CXCR、CD44、CD147的表达水平；同时将检测的细胞悬液,在荧光显微镜下观察细胞的荧光标记情况拍照记录；并将与其对应的垂体腺瘤石蜡标本,应用免疫组化的方法,研究CXCL12/CXCR4、CD44和CD147的表达情况；采用SPSS17.0统计处理。
结果依据内分泌临床表现、影像学检查、术中所见和病理诊断结果,将垂体腺瘤(35例)分为21例侵袭性垂体腺瘤和14例非侵袭性垂体腺瘤两个实验组。比较两组间肿瘤标记物的表达水平,流式细胞仪检测结果中CXCR4、CD44、CD147的P值分别为0.002、0.221和0.178,免疫组化结果中CXCR4、CXCL12、CD44、CD147的P值分别为0.04、0.01、0.516和0.855；垂体腺瘤中CXCR4/CXCL12的表达水平水平明显高于正常垂体；荧光显微镜观察显示肿瘤细胞部分表达所标记的肿瘤标记物。侵袭鞍底的垂体腺瘤CXCR4表达率明显高于侵袭海绵窦组(mean分别为58.3%和23.6%,P<0.01)。CXCR4/CXCL12、CD44、CD147与患者性别、年龄、肿瘤大小及功能无明显关系(P>0.05)。
结论流式细胞仪检测肿瘤标记物在垂体腺瘤细胞中的表达率上要比免疫组化准确、敏感；在侵袭性垂体腺瘤中CXCR4/CXCL12的表达水平显著升高,CD44、CD147的表达水平无明显差异。CXCR4/CXCL12可能通过ERK1/2MAP激酶级联反应和Pyk2的活化,在垂体腺瘤的侵袭性发生过程中起作用,可作为研究垂体腺瘤不良生物学行为的新的潜在标记物。
Objective Detect the expression levels of CXCR4, CXCL12, CD44, CD147in invasive and non-invasive pituitary adenomas, and comparatively analyze the relevance of these proteins and the invasiveness of pituitary adenoma and the probable mechanisms.
Materials and Methods Collected the fresh specimens of surgical resection of35pituitary adenoms of our neurosurgery department from January to April,2009; made them to single cell suspension, and detected the expression of CXCR4, CD44, CD147by flow cytometry; at the same time, put the suspension under fluorescence microscope, observed the fluorescent-labeled cells and took photographed records; used the immunohistochemical methods to research the expression of CXCR4, CXCL12, CD44, CD147in the corresponding paraffin specimens of pituitary adenoma, and photographed them; used the SPSS17.0for statistical analysis, and took P<0.05as statistically significant standard.
Results Based on clinical manifestations, imaging examinations, surgery and pathological diagnosis, the pituitary adenomas (35cases) were divided into invasive pituitary adenomas (21cases) and non-invasive pituitary adenomas (14cases) compairing Comparing the expression levels of these two groups, the P values of CXCR4, CD44and CD147were0.002,0.221and0.178respectively in the results of flow cytometry; the P values of CXCR4, CXCL12, CD44, CD147in immunohistochemistry method were0.04,0.01,0.516and0.855respectively; the expression levels of CXCR4/CXCL12in pituitary adenomas were higher than the levels in normal pituitary; fluorescence microscope showed that part of pituitary adenomas expressd the labelled tumor markers. The expression of CXCR4in pituitary adenoma which invaded buttom of sella turcica was significantly higher than that cavernous sinus invaded group(means were58.3%and23.6%, P<0.01). CXCR4/CXCL12, CD44, CD147had no significant relationship with patient gender, age, tumor size and function(P>0.05).
Conclusion Flow cytometry is more accurate and sensitive in the detection of expression rate of tumor markers in tumor cells than immunohistochemistry.In invasive pituitary pituitary adenomas the expressions of CXCR4/CXCL12are significant increased, the expressions of CD44, CD147has no significant difference. CXCR4/CXCL12play an important role in the process of invasiveness of pituitary adenoma, possibly though the ERK1/2MAP kinase cascade and Pyk2activation and can be a new potential marker for the study of the abnormal biological behaviors of pituitary adenoma.
引文
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25. Kurosaki M, Saegert W, Abe T. Expression of vascular endothelial growth factor in growth hormone -secreting pituitary adenomas:special reference to the octreotide treatment. Neurol Res.2008 Jun;30(5):518-22.
26. Korsisaari N, Ross J, Wu X,et al. Blocking vascular endothelial growth factor-A inhibits the growth of pituitary adenomas and lowers serum prolactin level in a mouse model of multiple endocrine neoplasia type 1. Clin Cancer Res.2008 Jan 1;14(1):249-58.
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1.黄垂学,侯永宏,刘运生,等.垂体腺瘤中Bcl-2和Bax的表达与其侵袭性的关系[J].中国医学工程,2007,15(3)：254-257.
2. JIALIANG WANG, HUI WANG, ZHIZHONG LI, et al. C-Myc Is Required for Maintenance of Glioma Cancer Stem Cells[J]. PLoS ONE,2008,3(11):1-11.
3. IKEDA H, YOSHIMOTO T. The relationship between c-myc protein expression, the bromodeoxyuridine laeling index and the biological behavior of pituitary adenomas[J]. Acta Neuropathol,1992,83:361-364.
4.范月超,万峰,张洪涛,等.抑制C-myc、整合素β1对侵袭性垂体腺瘤侵袭力影响的研究[J].脑与神经疾病杂,2005,13(5)：359-361.
5. JUNK DJ, VRBA L, WATTS GS, et al. Different Mutant Wild-Type p53 Combinations Cause a Spectrum of Increased Invasive Potential in Nonmalignant Immortalized Human Mammary Epithelial Cells[J]. Neoplasia,2008,10(5):450-461.
6. YOSHINORI T, LONG JIN, SCHEITHAUER BW, et al. P53 Gene Mutations in Pituitary Carcinomas[J]. Endocr Pathol,2007,18:217-222.
7. KUMAR K, MACAULAY RJ, KELLY M, et al. Absent p53 immunohistochemical staining in a pituitary carcinoma.Can J Neurol Sci,2001,28(2):174-178.
8. MACHIAVELLI G, COTIGNOLA J, DANILOWICZ K, et al. Expression of p16INK4A gene in human pituitary tumours[J]. Pituitary,2008,11:71-75.
9. TAKINO H, HERMAN V, WEISS M, et al. Purine-binding factor(nm23) gene expression in pituitary tumors:Marker of adenoma invasiveness.J Clin Endocrinol Metab,1995,80(5):1733-1738.
10. MUSAT M, KORBONITS M, KOLA B, et al. EnhaNFed protein kinase B/Akt signaling in pituitary tumours[J].Endocr Relat Cancer,2005,12(2):423-433.
11. ZHANG X, HORWITZ GA, HEANEY AP, et al. Pituitary tumor transforming gene(PTTG)expression in pituitary adenomas [J].J Clin Endocrinol Metab,1999,84(2):761-767.
12. MALIK MT, KAKAR SS. Regulation of angiogenesis and invasion by human Pituitary tumor transforming gene (PTTG) through iNFreased expression and secretion of matrix metalloproteinase-2 (MMP-2)[J]. Molecular Cancer,2006,5:61.
13. MINEMATSU T, SUZUKI M, SANNO N, et al. PTTG Overexpression Is Correlated with Angiogenesis in Human Pituitary Adenomas[J]. Endocrine Pathology,2006,11(2):143-154.
14. LEMAOULT J, KRAWICE-RADANNE I, DAUSSET J, et al. HLA-G1 expressing antigen-presenting cells induce immunosupp ressive CD4+T cells. Proc Natl Acad Sci USA,2004,101(18):7064-7069.
15.贾栋,高国栋.HLA-G蛋白在垂体瘤中的表达及意义[J].现代肿瘤医学,2007,15(9)：1245-1247.
16.石全红,霍钢,郑履平.Survivin在侵袭性垂体腺瘤中的表达及其与bcl-2、bax表达的关系[J].江西医学院学报,2005,45(4)：59-62.
17. HAUSER HP, BARDROFF M, PYROWOLAKIS G, et al. A giant ubiquitin conjugating enzyme related to IAP apoptosis inhibitors[J].J Cell Biol,1998,141:1415-1422.
18. OLIE RA, SIMOES-WUST AP, BAUMANN B. A novel antisense oligonucleotide targeting survivin expression induces apoptosis and sensitizes lung cancer cells to chemotherapy. Cancer Res,2000,60(11):2805-2809.
19. CHUI-XUE HUANG, YONG-HONG HOU, YUN-SHENG LIU. Expression of galectin-3 correlates with apoptosis in pituitary adenoma cells[J]. Neurosci Bull, 2008,24(1):34-38.
20. NAM DH, SONG SY, PARK K, et al.Clinical significance of molecular genetic changes in sporadic invasive pituitary adenomas. Exp Mol Med,2001, 33(3):111-116.
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